Journal
JOURNAL OF BIOTECHNOLOGY
Volume 301, Issue -, Pages 24-34Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jbiotec.2019.05.010
Keywords
Vitamin C; Gluconobacter oxydans; Enzymes; 2-Keto-L-gulonic acid; L-Ascorbic acid
Categories
Funding
- National Key Research and Development Program of China [2017YFC1600403]
- National Natural Science Foundation of China [31830068, 21822806]
- Fundamental Research Funds for the Central Universities [JUSRP51701A]
- National First-class Discipline Program of Light Industry Technology and Engineering [LITE2018-08]
- Distinguished Professor Project of Jiangsu Province
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2-Keto-L-gulonic acid (2-KLG) is the direct precursor of vitamin C in industrial synthesis. 2-KLG is mainly produced via the classical two-step fermentation route. In the two-step fermentation process, 2-KLG can be synthesized from L-sorbose by Ketogulonicigenium vulgare aided by Bacillus megaterium. There are five sorbose/sorbosone dehydrogenases (SSDHs), SSDA1, SSDA1-P, SSDA2, SSDA3 and SSDB, and two sorbosone dehydrogenases (SNDHs), glucose/sorbosone dehydrogenase (GSNDH) and sorbosone dehydrogenase (SNDH), in K. vulgare, which could play crucial roles in transforming L-sorbose or L-sorbosone to 2-KLG. However, confusion about the catalytic characteristics of the individual SSDHs and SNDHs makes construction of a recombinational strain for the purpose of enhancing 2-KLG production difficult. In this study, the five SSDHs and two SNDHs from K. vulgare WSH-001 were purified, and their optimal pH values and reaction temperatures, kinetic properties, thermostabilities, substrate spectra and effects of electron acceptors on their performances were systematically determined. Among these dehydrogenases, only SSDA1 and SSDA3 have high activity for catalyzing L-sorbose to 2-KLG directly. These data provide more clues for ways to achieve enhanced conversion of L-sorbose in K. vulgare, which could facilitate both the construction of a more efficient one-step fermentation 2-KLG producer and the reconstruction of a one-step fermentation process.
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